Screening for Peptides to Bind and Functionally Inhibit SARS-CoV-2 Fusion Peptide Using Mirrored Combinatorial Phage...

Area of Science:

• Virology
• Biochemistry
• Drug Discovery

Background:

• The conserved spike fusion peptide (FP) of coronaviruses presents a potential target for broad-spectrum antiviral therapies.
• Developing pancoronaviral inhibitors requires identifying molecules that can disrupt conserved viral mechanisms.

Purpose of the Study:

• To identify novel peptide inhibitors targeting the conserved SARS-CoV-2 spike fusion peptide (FP).
• To explore phage display libraries for discovering peptides with potential pancoronaviral inhibitory activity.

Main Methods:

• Screening of a random 7-mer peptide library (NEB PhD-7-mer) against the SARS-CoV-2 FP.
• Synthesis of identified peptides as proteolytically resistant D-peptides.
• Screening of a proteomic-derived phage display library from human disordered regions.
• Molecular dynamics structural modeling to elucidate binding interactions.

Main Results:

• Ten D-peptides from the random library showed no inhibition of SARS-CoV-2 infection in cell culture.
• Two overlapping 14mer peptides from OTUD1 were identified from the proteomic library.
• A synthetic peptide based on OTUD1 sequences did not significantly inhibit viral entry.
• Molecular modeling revealed a stable binding mode between an OTUD1 peptide and the FP, suggesting potential for future inhibitor design.

Conclusions:

• Phage display strategies, while not yielding direct inhibitors, facilitated the development of a computational model for FP disruptors.
• The study provides a foundation for designing future pancoronaviral FP inhibitors based on structural insights.